Difference between revisions of "Part:BBa K1321366:Design"

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===Design Notes===
 
===Design Notes===
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This part was made from the composite parts that we submitted, and fused by Biobrick and Freiburg cloning. SfGFP was fused as a reporter for our CBD binding assay, and may have uses for future teams as an easily quantifiable reporter.
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Codon optimised for E.coli.
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Once the individual parts were confirmed in Freiburg format, we cloned them using the AgeI and NgoMIV site, creating a scar that also functions as a small linker.
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The T7 vector was fused to the protein fusion using the Xba and SpeI site.
  
 
===Source===
 
===Source===

Latest revision as of 23:42, 21 October 2014

CBDcex fused to sfGFP with T7 promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 53
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 401


Design Notes

Codon optimised for E.coli. Once the individual parts were confirmed in Freiburg format, we cloned them using the AgeI and NgoMIV site, creating a scar that also functions as a small linker. The T7 vector was fused to the protein fusion using the Xba and SpeI site.

Source

CBDcex comes from Cellulomonas fimi, and sfGFP is a modified super-folding mutant of GFP. Our sfGFP part was an improvement of BBa_I746909, wherein Freiburg sites were added. Please see BBa_K1321337 for more info on this part. CBDcex was requested from the registry, as it was submitted by the Freiburg team as BBa_K863101.

The T7 vector was made by inverse PCR of BBa_I746909, and ligated. Please see our BBa_ K1321338 for more information on this part.

References